Atomic detail of chemical transformation at the transition state of an enzymatic reaction.
Transition path sampling (TPS) has been applied to the chemical step of human purine nucleoside phosphorylase (PNP). The transition path ensemble provides insight into the detailed mechanistic dynamics and atomic motion involved in transition state passage. The reaction mechanism involves early loss of the ribosidic bond to form a transition state with substantial ribooxacarbenium ion character, followed by dynamic motion from the enzyme and a conformational change in the ribosyl group leading to migration of the anomeric carbon toward phosphate, to form the product ribose 1-phosphate. Calculations of the commitment probability along reactive paths demonstrated the presence of a broad energy barrier at the transition state. TPS identified (i) compression of the O4'...O5' vibrational motion, (ii) optimized leaving group interactions, and (iii) activation of the phosphate nucleophile as the reaction proceeds through the transition state region. Dynamic motions on the femtosecond timescale provide the simultaneous optimization of these effects and coincide with transition state formation.
Saen-Oon
Suwipa
S
Departments of Biophysics and Biochemistry, Albert Einstein College of Medicine, 1300 Morris Park Ave, Bronx, NY 10461, USA.
Quaytman-Machleder
Sara
S
Schramm
Vern L
VL
Schwartz
Steven D
SD
D002384:Catalysis; D006801:Humans; D007700:Kinetics; D008956:Models, Chemical; D009038:Motion; D011336:Probability; D011487:Protein Conformation; D011683:Purine-Nucleoside Phosphorylase / Q000737:chemistry*; D012267:Ribosemonophosphates / Q000096:biosynthesis; D014732:Vibration
D012267:Ribosemonophosphates; C031154:ribose 1-phosphate; D011683:Purine-Nucleoside Phosphorylase